Fig. 2: Enhanced sister cell heterogeneity after replication stress.

a, A representative single-cell lineage from untreated 53BP1–PCNA U-2 OS cells. PCNA foci, 53BP1 foci and the area of the nucleus over 55 h of live imaging are shown. Sequential staining intensities from individual cells of the lineage (sc) and the mean intensities from the reference population (rp) are depicted for the markers pRb, γH2AX, p21 and p53. b, Representative video stills from the single-cell lineage in a and corresponding images from the sequential staining of cells for pRb, γH2AX, p21 and p53. Scale bar, 10 µm. c, Single-cell lineage from 53BP1–PCNA U-2 OS cells treated with 0.2 µM APH during the G2 phase of the cell cycle. PCNA foci, 53BP1 foci and area of the nucleus over 55 h of live imaging are shown. Sequential staining intensities from individual cells of the lineage and mean intensities from the corresponding reference populations are shown. UT, untreated. d, Single-cell lineage from 53BP1–PCNA U-2 OS cells treated with 1 µM of ATRi during the G2 phase of the cell cycle. PCNA foci, 53BP1 foci and the area of the nucleus over 55 h of live imaging are shown. Sequential staining intensities from individual cells of the lineage and mean intensities from the corresponding reference populations are shown. For the box plots, the box limits show the interquartile range (IQR, 25th percentile (Q1) to 75th percentile (Q3)), the median (centre line) and the whiskers define the lower and upper adjacent value; the dots show outliers smaller than Q1 − 1.5 × IQR and greater than Q3 + 1.5 × IQR; n > 35 end-point measurements per sample. Drugs in c and d were removed after 24 h.